大鼠肝再生的肝细胞凋亡中Kruppel样因子4 mRNA、微小RNA-881-3p、环状RNA_20298和环状RNA_14826的表达变化与作用

doi:10.16098/j.issn.0529-1356.2023.04.007

解剖学报 ›› 2023, Vol. 54 ›› Issue (4): 420-424.doi: 10.16098/j.issn.0529-1356.2023.04.007

大鼠肝再生的肝细胞凋亡中Kruppel样因子4 mRNA、微小RNA-881-3p、环状RNA_20298和环状RNA_14826的表达变化与作用

林凯琳^1,2杨献光^1,2王子慧^1,2 臧夏炎^1,2 薛奇杰^1,2韩璐^1,2张春博^1,2 赵志虎^3* 徐存拴^1,2*

1.河南师范大学生命科学学院，河南新乡 453007; 2.河南省-科技部共建细胞分化调控国家重点实验室培育基地，河南新乡 453007; 3.军事医学研究院生物工程研究所，北京 100071

收稿日期:2022-02-23 修回日期:2022-09-02 出版日期:2023-08-06 发布日期:2023-08-06
通讯作者: 徐存拴 E-mail:cellkeylab@126.com

Expression change and role of Kruppel-like factor 4 mRNA, microRNA-881-3p, circular RNA_20298 and circular RNA_14826 in the hepatocyte apoptosis during the rat liver regeneration

LIN Kai-lin^1,2 YANG Xian-guang^1,2 WANG Zi-hui^1,2 ZANG Xia-yan^1,2 XUE Qi-jie^1,2 HAN Lu^1,2 ZHANG Chun-bo^1,2 ZHAO Zhi-hu^3* XU Cun-shuan^1,2*

1.College of Life Science, He’nan Normal University, He’nan Xinxiang 453007, China; 2.State Key Laboratory Cultivation Base for Cell Differentiation Regulation, He’nan Xinxiang 453007, China; 3. Institute of Biotechnology，Academy of Military Medical Sciences, Beijing 10071，China

Received:2022-02-23 Revised:2022-09-02 Online:2023-08-06 Published:2023-08-06
Contact: 赵志虎;徐存拴 E-mail:cellkeylab@126.com

摘要/Abstract

摘要：

目的了解大鼠肝再生（LR）0 h和120 h时Kruppel样因子4（KLF4）基因及其mRNA相互作用的微小RNA（miRNA）和环状RNA（circRNA）的表达变化、相互作用和调节肝细胞凋亡的途径与方式。方法按Higgins等方法制备大鼠2/3肝切除（PH）模型，按Smedsrod等方法分离肝细胞，用大规模定量检测技术测定大鼠肝再生中肝细胞的mRNA、miRNA和circRNA合称内源竞争RNA（ceRNA）表达变化，用Cytoscape 3.2软件构建ceRNA的相互作用网络，用ceRNA综合分析等方法解析它们的表达相关性和作用相关性。结果 PH后0 h和120 h时，KLF4 mRNA的比值为1.00±0.16和3.14±0.27，miR.881.3p为18.30±1.44和0.47±0.02，circRNA_20298为0.32±0.10和4.24±0.22，circRNA_14826为0.42±0.13和0.61±0.08。同时，PH后0 h时，KLF4促进的肾上腺素受体β 2（ADRB2）、二甲基精氨酸二甲基氨基水解酶 2（DDAH2）、膜联蛋白A5（ANXA5）等4个细胞凋亡相关基因表达下调，抑制的细胞凋亡相关基因突触核蛋白γ（synuclein gamma，SNCG）、谷胱甘肽二硫化物还原酶（GSR）、FYVE、RhoGEF和PH结构域包含 4（FGD4）表达上调。PH后120 h时，KLF4促进的细胞凋亡相关基因ANXA5和胸腺素beta 10（TMSB10）表达上调，抑制的细胞凋亡相关基因氯化物细胞内通道4（CLIC4）和紫杉素3（ATXN3）表达下调。结论上述circRNA抑制的miRNA、miRNA抑制的KLF4 mRNA以及KLF4调节的细胞凋亡相关基因的表达相关性和作用相关性有利于PH后0 h时肝细胞保持活性状态和PH后120 h时肝细胞处于凋亡状态。

关键词: 肝再生, Kruppel样因子4, 生物高通量检测, 内源竞争RNA综合分析, 大鼠

Abstract:

Objective To explore the role pathway and pattern of the Kruppel-like factor 4 (KLF4) and its mRNA interaction with microRNA(miRNAs) and circular RNA(circRNAs) at 0 hour and the 120 th hour in the rat liver regeneration. Methods The rat 2/3 hepatectomy (partial hepatectomy, PH) model was prepared as described by Higgins, the hepatocytes were isolated according to the method of Smedsrod et al, the expression changes of mRNA, miRNA and circRNA together named as competing endogenous RNA (ceRNA) were detected by the large-scale quantitative detection technology, the interaction network of ceRNA was constructed by Cytoscape 3.2 software, and their correlation in expression and role were analyzed by ceRNA comprehensive analysis. Results It was found that at the 0 hour and the 120th hour PH, the ratio value of KLF4 mRNA showed 1.00±0.16 and 3.14±0.27, miR-881-3p displays 18.30±1.44 and 0.47±0.02, circRNA_20298 indicated 0.32±0.10 and 4.24±0.22, circRNA_14826 showed 0.42±0.13 and 0.61±0.08. At the same time, the four kinds of cell apoptosis-related genes adrenoceptor beta 2 (ADRB2), dimethylarginine dimethylaminohydrolase 2 (DDAH2), annexin A5 (ANXA5), ect, which were promoted in expression by KLF4, were down-regulated at 0 hour after PH, but the cell apoptosis-related genes synuclein gamma (SNCG), glutathione-disulfide reductase (GSR), FYVE, RhoGEF and PH domain containing 4 (FGD4), ect, which were inhibited in expression by KLF4, were upregulated at 0 hour after PH. On the other hand, the cell apoptosis-related genes ANXA5 and thymosin beta 10 (TMSB10), which are promoted in expression by KLF4, were up-regulated at the 120th hour after PH, but the cell apoptosis-related genes chloride intracellular channel 4 (CLIC4) and ataxin 3 (ATXN3), ect, which were inhibited in expression by KLF4, were down-regulated at the 120th hour after PH. Conclusion The correlation in expression and role of the miRNAs, which are inhibited by circRNAs, KLF4, its mRNA is inhibited by miRNAs, and the cell apoptosis-related genes, which are regulated by KLF4, are helpful for the hepatocyte to be in active state 0 hour after PH and to be in apoptotic state 120-hour after PH.

Key words: Liver regeneration, Kruppel like factor 4, Biological high-throughput detection, Competing endogenous RNA comprehensive analysis, Rat

中图分类号:

Q257

林凯琳杨献光王子慧臧夏炎薛奇杰韩璐张春博赵志虎徐存拴. 大鼠肝再生的肝细胞凋亡中Kruppel样因子4 mRNA、微小RNA-881-3p、环状RNA_20298和环状RNA_14826的表达变化与作用[J]. 解剖学报, 2023, 54(4): 420-424.

LIN Kai-lin YANG Xian-guang WANG Zi-hui ZANG Xia-yan XUE Qi-jie HAN Lu ZHANG Chun-bo ZHAO Zhi-hu XU Cun-shuan. Expression change and role of Kruppel-like factor 4 mRNA, microRNA-881-3p, circular RNA_20298 and circular RNA_14826 in the hepatocyte apoptosis during the rat liver regeneration [J]. Acta Anatomica Sinica, 2023, 54(4): 420-424.

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大鼠肝再生的肝细胞凋亡中Kruppel样因子4 mRNA、微小RNA-881-3p、环状RNA_20298和环状RNA_14826的表达变化与作用

Expression change and role of Kruppel-like factor 4 mRNA, microRNA-881-3p, circular RNA_20298 and circular RNA_14826 in the hepatocyte apoptosis during the rat liver regeneration

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